Guangdong Provincial Engineering Technology Research Center for Molecular Diagnosis and Innovative Drugs Translation of Cardiopulmonary Vascular Diseases, University Joint Laboratory of Guangdong Province and Macao Region on Molecular Targets and Intervention of Cardiovascular Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, China.
Department of Precision Laboratory, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, China.
BMC Infect Dis. 2024 Mar 18;24(1):326. doi: 10.1186/s12879-024-09212-4.
Currently, culture methods are commonly used in clinical tests to detect pathogenic fungi including Candida spp. Nonetheless, these methods are cumbersome and time-consuming, thereby leading to considerable difficulties in diagnosis of pathogenic fungal infections, especially in situations that respiratory samples such as alveolar lavage fluid and pleural fluid contain extremely small amounts of microorganisms. The aim of this study was to elucidate the utility and practicality of microfluidic chip technology in quick detection of respiratory pathogenic fungi.
DNAs of clinical samples (mainly derived from sputa, alveolar lavage fluid, and pleural fluid) from 64 coastal patients were quickly detected using microfluidic chip technology with 20 species of fungal spectrum and then validated by Real-time qPCR, and their clinical baseline data were analyzed.
Microfluidic chip results showed that 36 cases infected with Candida spp. and 27 cases tested negative for fungi, which was consistent with Real-time qPCR validation. In contrast, only 16 cases of fungal infections were detected by the culture method; however, one of the culture-positive samples tested negative by microfluidic chip and qPCR validation. Moreover, we found that the patients with Candida infections had significantly higher rates of platelet count reduction than fungi-negative controls. When compared with the patients infected with C. albicans alone, the proportion of males in the patients co-infected with multiple Candidas significantly increased, while their platelet counts significantly decreased.
These findings suggest that constant temperature amplification-based microfluidic chip technology combined with routine blood tests can increase the detection speed and accuracy (including sensitivity and specificity) of identifying respiratory pathogenic fungi.
目前，临床检测常采用培养方法来检测包括念珠菌属（Candida spp.）在内的致病真菌。然而，这些方法繁琐且耗时，因此在诊断致病真菌感染方面存在相当大的困难，尤其是在肺泡灌洗液和胸腔积液等呼吸道样本中微生物含量极少的情况下。本研究旨在阐明微流控芯片技术在快速检测呼吸道致病真菌方面的实用性。
使用微流控芯片技术对 64 例沿海地区患者的临床样本（主要来源于痰液、肺泡灌洗液和胸腔积液）的 DNA 进行快速检测，检测涵盖 20 种真菌谱，并通过实时 qPCR 进行验证，同时分析其临床基线数据。
微流控芯片结果显示，36 例感染念珠菌属，27 例真菌检测阴性，与实时 qPCR 验证结果一致。相比之下，培养方法仅检测到 16 例真菌感染；然而，有一个培养阳性样本经微流控芯片和 qPCR 验证为阴性。此外，我们发现念珠菌感染者的血小板计数减少率明显高于真菌阴性对照组。与单独感染白色念珠菌的患者相比，合并多种念珠菌感染的患者中男性比例明显增加，而血小板计数明显降低。
这些发现表明，基于恒温扩增的微流控芯片技术与常规血液检查相结合，可以提高识别呼吸道致病真菌的检测速度和准确性（包括灵敏度和特异性）。
